张 松1，黄万兵2，刘宏宇2，朱国胜2，尚念杰2，桂 阳2*，赵德刚1*
（1.贵州大学生命科学学院/农业生物工程研究院，山地植物资源保护与保护种质创新教育部重点实验室，山地生态与农业生物工程协同创新中心，贵州贵阳 550025
2.贵州省农业科学院农作物品种资源研究所，贵州贵阳 550006）
摘要：为探究冬荪液体深层培养不同时期代谢物动态变化及其关系，基于液体深层培养技术，超高效液相色谱-串联飞行时间质谱联用技术研究了不同发酵时间菌丝体分泌物差异情况。结果表明，534种差异代谢物隶属于有机酸及衍生物、有机氧化合物、苯丙素和聚酮类化合物等13个不同化学类别，有机酸类化合物占比为17.978%是发酵环境成酸性的主要原因。通过主成分分析和正交偏最小二乘判别分析，以FC＞1.5或FC＜0.67，P value＜0.05为条件筛选鉴定到大豆苷，大豆苷元，水苏糖，反式-4-羟基-L-脯氨酸，L-二氢月桂酸盐，UDP-GlcNAc等37种特征代谢物。KEGG通路分析表明，差异代谢物显著富集在ABC转运蛋白、cAMP信号通路、半乳糖代谢等86条代谢途径，推测了冬荪水苏糖等胞外多糖的合成路径，对鬼笔属真菌胞外多糖研究具有指导意义。不同发酵时间代谢物种类和数量远高于已研究报道的子实体和菌丝体化学成分，为今后深层研究冬荪活性成分提供理论参考。
关键词：药用真菌；发酵液；超高效液相色谱-串联质谱；差异代谢物；水苏糖
中图分类号：TS219 文献标识码：A 文章编号：1674-506X（2021）02-0014-0010
Study on Extracellular Metabolites of Phallus dongsun in Different Periods of Submerged Fermentation
ZHANG Song1, HUANG Wan-bing2, LIU Hong-yu2, ZHU Guo-sheng2, SHANG Nian-jie2, GUI Yang2*, ZHAO De-gang1*
（1.Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region （Ministry of
Education）, Collaborative Innovation Center for Mountain Ecology & Agro-Bioengineering, College of Life Sciences/Institute
of Agro-bioengineering, Guizhou University, Guiyang Guizhou 550025, China;
2.Guizhou Institute of Crop Germplasm Resources, Guizhou Academy of Agricultural Sciences, Guiyang Guizhou 550006, China）
Abstract：In order to explore the dynamic changes and relationship of metabolites in mycelia fermentation culture of Phallus dongsun, use submerged fermentation technology and UHPLC-Q-TOF MS to study the difference of mycelium secretion in different fermentation time. Research shows that the 534 metabolites belong to 13 different chemical categories, including organic acids and derivatives, organic oxygen compounds, phenylpropanoids and polyketides. 17.978% of organic acids are the main reason for the acidity of the fermentation environment. PCA and OPLS- DA with FC＞1.5 or FC＜0.67, P value＜0.05 showed that 37 characteristic metabolites were screened and identified, such as daiazin, daidzein, stachyose, trans-4-hydroxy-L-proline, L-dihydrolaurate, UDP- GlcNAc. Differential metabolites are significantly enriched in 86 metabolic pathways, including ABC transporter, cAMP signaling pathway, galactose metabolism and the synthetic pathway of exopolysaccharides such as stachyose was speculated. It has guiding significance for the study of extracellular polysaccharides ofPhallus. The type and quantity of metabolites in different fermentation time were much higher than the chemical composition of fruiting body and mycelium, which provided a theoretical reference for the further study of active components in Phallus dongsun.
Keywords：medicinal fungi; fermentation broth; UHPLC-MS/MS; differential metabolite; stachyose
doi：10.3969/j.issn.1674-506X.2021.02-002
收稿日期：2021-01-22
基金项目：贵州省菌物资源创新利用（黔科合支撑［2019］2451-6）；贵州省药用菌产业化配套技术集成服务企业行动计划（黔科合服企［2018］4006）；贵州省高层次创新型人才培养（黔科合平台人才［2016］4013）；冬荪菌种选育及扩繁关键技术研究与应用示范（黔科合重大专项［2019］3008-2号）；贵州省食用菌育种重点实验室（黔科合平台人才［2019］5105）；贵州省天麻竹荪育种和栽培科技创新人才团队（黔科合平台人才［2018］5619）
作者简介：张松（1994-），男，硕士研究生。研究方向：生化与分子。
*通信作者
引文格式：张松,黄万兵,刘宏宇,等.冬荪液体深层培养不同时期胞外代谢物研究[J].食品与发酵科技,2021,57(2):14-22,29.
